Presence of mecA-positive multidrug-resistant Staphylococcus epidermidis in bovine milk samples in Brazil.

Bacteria of the genus Staphylococcus are one of the major pathogens causing bovine mastitis. In recent decades, resistance of this genus to oxacillin (methicillin) has been a matter of concern due to the possibility of reducing the effectiveness of mastitis treatments and the transfer of resistance determinants to other bacteria. Oxacillin resistance was studied in 170 staphylococci from bovine milk samples, including 79 Staphylococcus aureus and 91 coagulase-negative staphylococci (CNS). The susceptibility profile of 10 antimicrobial agents used in veterinary practice was determined by the Etest method. In addition to the Etest, the phenotypic characterization of oxacillin resistance was tested using the cefoxitin disk diffusion test. All isolates were screened by PCR to detect the mecA gene in 2 different regions of the gene. The isolates with an oxacillin minimum inhibitory concentration ≥0.5 µg/mL or resistant to cefoxitin were identified by sequencing a 536-bp fragment of the 16S rRNA gene. This group of isolates was also evaluated for the presence of blaZ and mecC genes. Molecular analysis of the mecA gene was carried out by typing of the staphylococcal cassette chromosome mec (SCCmec). The relatedness of the mecA-positive isolates was evaluated by macrorestriction of chromosomal DNA followed by pulsed-field gel electrophoresis. With the exception of penicillin and oxacillin, 86% of the isolates showed susceptibility to cephalothin, gentamicin, erythromycin, sulfonamide, trimethoprim-sulfamethoxazole, and tetracycline. All S. aureus isolates were susceptible to oxacillin, whereas 47% (n=43) of the CNS isolates were resistant. The CNS isolates showed a higher resistance to cephalothin, erythromycin, tetracycline, and gentamicin in comparison with S. aureus. The mecA gene was only detected in 10 CNS isolates, identified as Staphylococcus epidermidis, and classified into 3 pulsotypes (A, B, and C) and 4 subtypes (A1, B1, B2, and B3). Among the isolates with an oxacillin resistance phenotype, 12 were positive for the blaZ gene, and 9 of them were mecA-positive. Two of the oxacillin-resistant isolates amplified the mecA homolog gene of Staphylococcus sciuri and none amplified mecC. Three SCCmec types, I, IV, and V, were found. Our results suggest that Staphylococcus epidermidis can be a reservoir for mecA for other Staphylococcus species. Studies investigating the molecular and phenotypic profile of antimicrobial resistance in staphylococcal species should be performed for controlling the spread of resistance and the selection of appropriate therapeutic measures.

Is Galleria mellonella model a good alternative to study virulence in Staphylococcus aureus from bovine mastitis?

Staphylococcus aureus is one of the agents of bovine mastitis of hardest control due to a complex pathogenesis comprising a variety of virulence factors, which ensures its persistence in the mammary gland, causing significant health and economic losses. Therefore, understanding the pathogenesis of this agent is imperative. Galleria mellonella has stood out as an invertebrate animal model for the study of infectious diseases that affect several hosts. This work aimed to evaluate G. mellonella larvae as an experimental model for the study of virulence phenotypes in an S. aureus population isolated from bovine mastitis. Thirty genetically divergent S. aureus strains were chosen based on PFGE analysis. After experimental infection, larvae survival rates, bacterial growth in hemolymph, melanization intensity of the dorsal vessel, and histological characteristics of the infected tissues were evaluated. The G. mellonella model showed a clear diversity in the S. aureus pathogenicity pattern, allowing the differentiation of strains with virulence phenotypes ranging from high to low degrees. Histological analysis confirmed that the strains tested were capable of inducing the formation of nodules and melanization spots in the dorsal vessels of the larvae in different magnitudes. The strains 16S-717, 19C-828, and 31S-1443 presented the highest virulence intensity among the bacteria tested and will be used further for the generation of S. aureus mutant populations to prospect genetic targets aimed to develop control strategies of bovine mastitis. Altogether, our results suggest that G. mellonella is an attractive and low-cost animal model for characterizing virulence phenotypes of large S. aureus populations.

Interference of Streptococcus agalactiae Blitz Therapy in Staphylococcus aureus Microbiological Diagnosis in Subclinical Bovine Mastitis.

Bovine mastitis is an important and costly disease to dairy cattle. Diagnostic methods usually performed in Brazil are somatic cell counts (SCC) and milk microbiology. Low bacteria shedding in milk implies no colony growth in microbiological tests and false negative results. Streptococcus agalactiae and Staphylococcus aureus are principal pathogens of mixed mastitis. However, S. agalactiae has a higher bacterial release from the mammary gland than S. aureus, affecting microbiological sensitivity to diagnose S. aureus. This study aimed to estimate the SCC and total bacterial count (TBC) from cows according to pathogen isolated in milk and to evaluate variation in S. aureus diagnosis by a microbiological test during S. agalactiae treatment, which is called blitz therapy. Both S. agalactiae and S. aureus presented high SCC means, although S. agalactiae showed shedding of bacteria 2.3 times greater than S. aureus. Microbiological sensitivity to S. aureus increased for 5 months during S. agalactiae treatment. The prevalence of S. agalactiae fell after 5 months of therapeutic procedures. The prevalence of S. aureus increased to 39.0. The results showed that due to high sensitivity, the polymerase chain reaction (PCR) could be used at the beginning of blitz therapy with the goal of S. agalactiae eradication from the dairy herd.

Recovery of Brucella in raw milk Minas artisanal cheese approved for consumption by official inspection agency in Brazil: assessment of prevalence and risk factors through One Health integrated approaches.

BACKGROUND: Minas artisanal cheese (MAC) from the Serro region is a Brazilian intangible cultural heritage. Produced from raw milk, it may carry zoonotic pathogens such as Brucella. This study included a randomized survey for the prevalence of Brucella-positive MAC and its associated factors. METHODS: MAC samples (n=55), each one from a different rural family-based cheese-processing agroindustry, were analysed for Brucella by direct polymerase chain reaction (PCR) species-specific DNA detection and cultivation-based approaches. RESULTS: Among 55 MACs that were analysed, we found 17 Brucella DNA-positive samples (30.9% [95% confidence interval {CI} 18.7 to 43.1]) by PCR and, for the first time, from one MAC (1.8% [95% CI 0.5 to 9.7]), viable Brucella abortus was recovered by cultivation. Higher values for two variables, the number of lactating cows per herd (p=0.043) and daily milk production per herd (p=0.043), were each associated with Brucella-positive MAC, which concentrated in three high-risk and one low-risk spatial clusters. CONCLUSIONS: MAC may be a source of Brucella for humans, since the positive samples were from batches that were sold by cheesemakers. This should be of concern and encourage cooperation between the health and agriculture sectors in order to mitigate this public health risk through One Health integrated approaches.

First molecular detection of Coxiella burnetii in Brazilian artisanal cheese: a neglected food safety hazard in ready-to-eat raw-milk product.

BACKGROUND: Global publications on Q fever have increased after the 2007 epidemic in the Netherlands. However, the epidemiology of Q fever/coxiellosis in Brazil is still poorly understood. Accordingly, there have been few studies investigating the presence of Coxiella burnetii in dairy products around the world, especially in Brazil, where consumption of fresh cheese made from raw-milk is very high. OBJECTIVE: This study was a random survey to assess the prevalence of C. burnetii by PCR in traditional Minas artisanal cheese from the Serro microregion, Brazil, which is manufactured from bovine raw-milk. METHODS: DNA extracted from 53 cheese samples were analyzed by nested PCR with C. burnetii-specific primers and the products confirmed by DNA sequencing. RESULTS: Out of the 53 cheese samples five (9.43%) were C. burnetii DNA-positive, each coming from one of the respective randomly selected manufacturing agroindustries. Based on our results, it is estimated that 1.62 tons/day of ready-to-eat cheese made from raw-milk from a total of 16.2 tons produced daily in the study region are contaminated with C. burnetii. CONCLUSION: To our knowledge, this is the first report of highly heat-resistant zoonotic pathogen in raw-milk Brazilian artisanal cheese. This food safety hazard has been completely neglected in ready-to-eat raw-milk Brazilian artisanal cheese and could imply potential threats to consumers, since C. burnetii survives in artisanal cheese submitted to long ripening periods. Thus, this work established random and representative baseline prevalence of C. burnetii in this food product in Brazil. Further epidemiological studies, monitoring trends and setting control targets are warranted. Finally, these results point out the importance of including C. burnetii in animal and public health surveillance programs.

Gel mobility shift scanning of pectin-inducible promoter from Penicillium griseoroseum reveals the involvement of a CCAAT element in the expression of a polygalacturonase gene.

Previous reports have described pgg2, a polygalacturonase-encoding gene of Penicillium griseoroseum, as an attractive model for transcriptional regulation studies, due to its high expression throughout several in vitro growth conditions, even in the presence of non-inducing sugars such as sucrose. A search for regulatory motifs in the 5' upstream regulatory sequence of pgg2 identified a putative CCAAT box that could justify this expression profile. This element, located 270 bp upstream of the translational start codon, was tested as binding target for regulatory proteins. Analysis of a 170 bp promoter fragment by electrophoretic mobility shift assay (EMSA) with nuclear extracts prepared from mycelia grown in pectin-containing culture medium revealed a high mobility complex that was subsequently confirmed by analyzing it with a double-stranded oligonucleotide spanning the CCAAT motif. A substitution in the core sequence for GTAGG partially abolished the formation of specific complexes, showing the involvement of the CCAAT box in the regulation of the polygalacturonase gene studied.

[Seasonality of dengue reporting in state capitals in the Brazilian Amazon and impacts of El Niño/La Niña]. / Sazonalidade nas notificações de dengue das capitais da Amazônia e os impactos do El Niño/La Niña.

The dynamics of dengue transmission are multifactorial and involve socioeconomic, ecological, and environmental aspects, the latter being closely related to local climatic conditions that affect the vector's reproductive cycle. Climate depends in turn on tropical oceanic mechanisms such as phases of El Niño/La Niña over the Pacific. The study contributes to this discussion and reports on the correlations between the Multivariate ENSO Index (MEI) in the Pacific and the number of reported dengue cases in seven state capitals in the Brazilian Amazon from 2001 to 2012. The study also analyzes the seasonality pattern (quarterly mean values) in dengue cases throughout the region. Evidence that El Niño/La Niña causes a decrease versus increase in the local rainfall pattern is consistent with the lower versus higher number of reported dengue cases in most of the state capitals in the Amazon, a result proven by the statistically significant negative correlations seen in Manaus (Amazonas), São Luís (Maranhão), Belém (Pará) and Palmas (Tocantins). The 12-years means (2001-2012) revealed the presence of pronounced seasonality in dengue incidence in the majority of the state capitals, with sharp peaks from January to March [Rio Branco (Acre), Manaus, Belém and Palmas] and from April to June (São Luís), corresponding to 50-70% of the annual total. State capitals farther north [Boa Vista (Roraima) and Macapá (Amapá)] showed dengue reporting in all quarters of the year, with no pronounced seasonality.

A dinâmica de transmissão da dengue é multifatorial e envolve aspectos socioeconômicos, ecológicos e ambientais, sendo este último intimamente relacionado às condições climáticas locais que interferem no ciclo reprodutivo dos vetores da doença. Por sua vez, o clima é dependente dos mecanismos oceânicos tropicais, a exemplo das fases de El Niño/La Niña sobre o Pacífico. O presente trabalho contribui com esta temática e reporta as correlações entre o índice MEI (Multivariate ENOS Index) do Pacífico e o número de casos notificados de dengue em sete capitais da Amazônia brasileira, no período de 2001 a 2012. Além disso, investiga-se o padrão de sazonalidade (médias trimestrais) dos casos de dengue ao longo da região. As evidências de que o fenômeno El Niño/La Niña provoca redução/aumento no padrão pluviométrico local é consistente com o número menor/maior de casos notificados de dengue na maior parte das capitais amazônicas, cujo resultado foi comprovado pelas correlações negativas estatisticamente significantes encontradas para Manaus (Amazonas), São Luís (Maranhão), Belém (Pará) e Palmas (Tocantins). As médias dos 12 anos (2001/2012) revelaram a presença de sazonalidade pronunciada na incidência de dengue na maioria das capitais, com picos acentuados de janeiro a março [Rio Branco (Acre), Manaus, Belém e Palmas] e de abril a junho (São Luís), correspondendo em torno de 50% a 70% do total anual. As localidades mais ao norte [Boa Vista (Roraima) e Macapá (Amapá)] revelaram registro da dengue ao longo de todos os trimestres do ano, não apresentando sazonalidade acentuada.

La dinámica de transmisión del dengue es multifactorial e implica aspectos socioeconómicos, ecológicos y ambientales, estando estos últimos íntimamente relacionados con las condiciones climáticas locales que interfieren en el ciclo reproductivo de los vectores de la enfermedad. A su vez, el clima depende de los mecanismos oceánicos tropicales, como por ejemplo las fases de El Niño/La Niña sobre el Pacífico. El presente trabajo contribuye a esta temática e informa sobre las correlaciones entre el índice MEI (Multivariate ENOS Index) del Pacífico y el número de casos notificados de dengue en siete capitales de la Amazonia brasileña durante el período de 2001 a 2012. Además, se investiga el patrón de estacionalidad (medias trimestrales) de los casos de dengue a lo largo de la región. Las evidencias de que el fenómeno El Niño/La Niña provoca reducción/aumento en el patrón pluviométrico local es consistente con el número menor/mayor de casos notificados de dengue en la mayor parte de las capitales amazónicas, cuyo resultado se comprobó por las correlaciones negativas estadísticamente significativas encontradas en Manaus (Amazonas), São Luís (Maranhão), Belém (Pará) y Palmas (Tocantins). Las medias de los doce años (2001/2012) revelaron la presencia de estacionalidad pronunciada en la incidencia de dengue en la mayoría de las capitales, con picos acentuados de enero a marzo [Rio Branco (Acre), Manaus, Belém y Palmas] y de abril a junio (São Luís), correspondiendo en torno del 50 al 70% del total anual. Las localidades más al norte [Boa Vista (Roraima) y Macapá (Amapá)] revelaron registro del dengue a lo largo de todos los trimestres del año, no presentando una estacionalidad acentuada.

First molecular detection of Coxiella burnetii in Brazilian artisanal cheese: a neglected food safety hazard in ready-to-eat raw-milk product

ABSTRACT Background: Global publications on Q fever have increased after the 2007 epidemic in the Netherlands. However, the epidemiology of Q fever/coxiellosis in Brazil is still poorly understood. Accordingly, there have been few studies investigating the presence of Coxiella burnetii in dairy products around the world, especially in Brazil, where consumption of fresh cheese made from raw-milk is very high. Objective: This study was a random survey to assess the prevalence of C. burnetii by PCR in traditional Minas artisanal cheese from the Serro microregion, Brazil, which is manufactured from bovine raw-milk. Methods: DNA extracted from 53 cheese samples were analyzed by nested PCR with C. burnetii-specific primers and the products confirmed by DNA sequencing. Results: Out of the 53 cheese samples five (9.43%) were C. burnetii DNA-positive, each coming from one of the respective randomly selected manufacturing agroindustries.Based on our results, it is estimated that 1.62 tons/day of ready-to-eat cheese made from raw-milk from a total of 16.2 tons produced daily in the study region are contaminated with C. burnetii. Conclusion: To our knowledge, this is the first report of highly heat-resistant zoonotic pathogen in raw-milk Brazilian artisanal cheese. This food safety hazard has been completely neglected in ready-to-eat raw-milk Brazilian artisanal cheese and could imply potential threats to consumers, since C. burnetii survives in artisanal cheese submitted to long ripening periods. Thus, this work established random and representative baseline prevalence of C. burnetii in this food product in Brazil. Further epidemiological studies, monitoring trends and setting control targets are warranted. Finally, these results point out the importance of including C. burnetii in animal and public health surveillance programs.

Seroprevalence for equine infectious anaemia in Equidae seized in the municipality of Petrópolis, State of Rio de Janeiro, Brazil, 2015/2018 / Soroprevalência para anemia infecciosa equina em Equídeos apreendidos no município de Petrópolis, RJ, Brasil, 2015/2018

ABSTRACT: Equine infectious anaemia (EIA) is a globally distributed disease caused by a lentivirus. EIA is a notifiable disease that should be reported to the Official Veterinary Service (SVO). Outbreaks are controlled by sacrificing the EIA-positive animals. The study analysed the prevalence of EIA in 165 Equidae seized on public roads in the municipality of Petrópolis, Rio de Janeiro, Brazil, from 2015 to 2018. After the seizure and identification of the Equidae, blood was collected to diagnose EIA. A prevalence of 11.8% was observed among the Equidae seized, and the main risk factor was the region of seizure. A kernel-smoothed map showed areas with the highest density of EIA-positive Equidae in the centre and to the east of the study area. In this region, the chance of seizing an equine positive for EIA was greater (OR=3.6) relative to the remaining region. The implementation and maintenance of Equidae seizures and subsequent diagnostic testing for EIA in the municipality of Petrópolis were shown to have great importance for the control and prevention of the disease. Indicating the place of seizure on the identification cards of the animals enabled delineation of the locations in the municipality of Petrópolis, that had a higher risk of EIA-positive animals.

RESUMO: A anemia infecciosa equina (AIE) é uma doença globalmente distribuída causada por um lentivírus. AIE é uma doença de notificação obrigatória que deve ser relatada ao Serviço Veterinário Oficial (SVO), e os surtos são controlados pelo sacrifício dos animais positivos para AIE. O presente estudo analisou a prevalência de AIE em 165 equídeos apreendidos em vias públicas no município de Petrópolis, estado do Rio de Janeiro, Brasil, no período de 2015 a 2018. Após a apreensão e identificação dos equídeos seu sangue foi coletado para diagnosticar a AIE. Observou-se uma prevalência real de 11,8% entre os equídeos apreendidos, sendo que o principal fator de risco identificado foi a região de apreensão. O Mapa de Kernel mostrou que as áreas com maior densidade de equídeos positivos estavam no centro e leste da área de estudo. Nesta região, houve maior chance (OR = 3,6) de apreender um animal positivo para AIE em relação ao da outra região, composto pelos 34 locais restantes. A implementação e manutenção de apreensão de equídeos e posterior teste de diagnóstico para AIE no município de Petrópolis, RJ, mostrou-se de grande importância para o controle e prevenção da doença. A indicação do local da apreensão nos cartões de identificação dos animais possibilitou um delineamento dos locais no município de Petrópolis, RJ, que apresentavam maior risco de animais positivos para AIE.

Sazonalidade nas notificações de dengue das capitais da Amazônia e os impactos do El Niño/La Niña / Seasonality of dengue reporting in state capitals in the Brazilian Amazon and impacts of El Niño/La Niña / Estacionalidad en las notificaciones de dengue de las capitales de la Amazonia y los impactos de El Niño/La Niña

Resumo: A dinâmica de transmissão da dengue é multifatorial e envolve aspectos socioeconômicos, ecológicos e ambientais, sendo este último intimamente relacionado às condições climáticas locais que interferem no ciclo reprodutivo dos vetores da doença. Por sua vez, o clima é dependente dos mecanismos oceânicos tropicais, a exemplo das fases de El Niño/La Niña sobre o Pacífico. O presente trabalho contribui com esta temática e reporta as correlações entre o índice MEI (Multivariate ENOS Index) do Pacífico e o número de casos notificados de dengue em sete capitais da Amazônia brasileira, no período de 2001 a 2012. Além disso, investiga-se o padrão de sazonalidade (médias trimestrais) dos casos de dengue ao longo da região. As evidências de que o fenômeno El Niño/La Niña provoca redução/aumento no padrão pluviométrico local é consistente com o número menor/maior de casos notificados de dengue na maior parte das capitais amazônicas, cujo resultado foi comprovado pelas correlações negativas estatisticamente significantes encontradas para Manaus (Amazonas), São Luís (Maranhão), Belém (Pará) e Palmas (Tocantins). As médias dos 12 anos (2001/2012) revelaram a presença de sazonalidade pronunciada na incidência de dengue na maioria das capitais, com picos acentuados de janeiro a março [Rio Branco (Acre), Manaus, Belém e Palmas] e de abril a junho (São Luís), correspondendo em torno de 50% a 70% do total anual. As localidades mais ao norte [Boa Vista (Roraima) e Macapá (Amapá)] revelaram registro da dengue ao longo de todos os trimestres do ano, não apresentando sazonalidade acentuada.

Abstract: The dynamics of dengue transmission are multifactorial and involve socioeconomic, ecological, and environmental aspects, the latter being closely related to local climatic conditions that affect the vector's reproductive cycle. Climate depends in turn on tropical oceanic mechanisms such as phases of El Niño/La Niña over the Pacific. The study contributes to this discussion and reports on the correlations between the Multivariate ENSO Index (MEI) in the Pacific and the number of reported dengue cases in seven state capitals in the Brazilian Amazon from 2001 to 2012. The study also analyzes the seasonality pattern (quarterly mean values) in dengue cases throughout the region. Evidence that El Niño/La Niña causes a decrease versus increase in the local rainfall pattern is consistent with the lower versus higher number of reported dengue cases in most of the state capitals in the Amazon, a result proven by the statistically significant negative correlations seen in Manaus (Amazonas), São Luís (Maranhão), Belém (Pará) and Palmas (Tocantins). The 12-years means (2001-2012) revealed the presence of pronounced seasonality in dengue incidence in the majority of the state capitals, with sharp peaks from January to March [Rio Branco (Acre), Manaus, Belém and Palmas] and from April to June (São Luís), corresponding to 50-70% of the annual total. State capitals farther north [Boa Vista (Roraima) and Macapá (Amapá)] showed dengue reporting in all quarters of the year, with no pronounced seasonality.

Resumen: La dinámica de transmisión del dengue es multifactorial e implica aspectos socioeconómicos, ecológicos y ambientales, estando estos últimos íntimamente relacionados con las condiciones climáticas locales que interfieren en el ciclo reproductivo de los vectores de la enfermedad. A su vez, el clima depende de los mecanismos oceánicos tropicales, como por ejemplo las fases de El Niño/La Niña sobre el Pacífico. El presente trabajo contribuye a esta temática e informa sobre las correlaciones entre el índice MEI (Multivariate ENOS Index) del Pacífico y el número de casos notificados de dengue en siete capitales de la Amazonia brasileña durante el período de 2001 a 2012. Además, se investiga el patrón de estacionalidad (medias trimestrales) de los casos de dengue a lo largo de la región. Las evidencias de que el fenómeno El Niño/La Niña provoca reducción/aumento en el patrón pluviométrico local es consistente con el número menor/mayor de casos notificados de dengue en la mayor parte de las capitales amazónicas, cuyo resultado se comprobó por las correlaciones negativas estadísticamente significativas encontradas en Manaus (Amazonas), São Luís (Maranhão), Belém (Pará) y Palmas (Tocantins). Las medias de los doce años (2001/2012) revelaron la presencia de estacionalidad pronunciada en la incidencia de dengue en la mayoría de las capitales, con picos acentuados de enero a marzo [Rio Branco (Acre), Manaus, Belém y Palmas] y de abril a junio (São Luís), correspondiendo en torno del 50 al 70% del total anual. Las localidades más al norte [Boa Vista (Roraima) y Macapá (Amapá)] revelaron registro del dengue a lo largo de todos los trimestres del año, no presentando una estacionalidad acentuada.

Overproduction of polygalacturonase by Penicillium griseoroseum recombinant strains and functional analysis by targeted disruption of the pgg2 gene.

Inactivation of the pgg2 gene, a polygalacturonase-encoding gene from Penicillium griseoroseum, reduced the total activity of polygalacturonase (PG) by 90 % in wild-type P. griseoroseum, which indicates that the pgg2 gene is the major gene responsible for PG production in this species. To increase PG production, the coding region of the pgg2 gene was cloned under the control of the glyceraldehyde 3-phosphate dehydrogenase (gpd) promoter and the terminator region of the tryptophan synthase (trpC) gene from Aspergillus nidulans (pAN52pgg2 vector). This vector was then used to transform P. griseoroseum. The transformed strains were characterized according to PG production using glucose, sucrose, or sugar cane juice as the carbon sources. The recombinant P. griseoroseum T146 strain contained an additional copy of the pgg2 gene, which resulted in a 12-fold increase in PG activity when compared with that detected in the supernatant of the control PG63 strain. The proteins secreted by the recombinant strain T146 showed a strong band at 38 kDa, which corresponds to the molecular weight of PG of the P. griseoroseum. The results demonstrate the significant biotechnological potential of recombinant P. griseoroseum T146 for use in PG production.

Sexuality and sickle cell anemia.

BACKGROUND: Sickle cell disease, the most common hereditary blood disease in the world, is the result of an atypical hemoglobin called S (Hb S) which, when homozygous (Hb SS) is the cause of sickle cell anemia. Changes of puberty, correlated with a delayed growth spurt, begin late in both male and female sickle cell anemia individuals with repercussions on sexuality and reproduction. The objectives of this exploratory and descriptive study were to characterize the development of sexuality in adults with sickle cell anemia by investigating the patient's perception of their sex life, as well as the information they had and needed on this subject. METHODS: Twenty male and female sickle cell anemia patients treated at the Hemocentro Regional de Uberaba (UFTM) with ages between 19 and 47 years old were enrolled. A socioeconomic questionnaire and a semi-structured interview on sexuality, reproduction and genetic counseling were applied. RESULTS: This study shows that the sickle cell anemia patients lacked information on sexuality especially about the risks of pregnancy and the possible inheritance of the disease by their children. Moreover, the sexual life of the patients was impaired due to pain as well as discrimination and negative feelings experienced in close relationships. CONCLUSION: The health care of sickle cell anemia patients should take into account not only the clinical aspects of the disease, but also psychosocial aspects by providing counseling on sexuality, reproduction and genetics, in order to give this population the possibility of a better quality of life.

Pain measurement as part of primary healthcare of adult patients with sickle cell disease.

OBJECTIVE: The aim of this exploratory, cross-sectional study was to evaluate pain in sickle cell disease patients and aspects related to primary healthcare. METHODS: Data were obtained through home interviews. The assessment instruments (body diagram, Numerical Pain Scale, McGill Pain Questionnaire) collected information on the underlying disease and on pain. Data were analyzed using the Statistical Package for Social Sciences program for Windows. Associations between the subgroups of sickle cell disease patients (hemoglobin SS, hemoglobin SC, sickle ß-thalassemia and others) and pain were analyzed using contingency tables and non-parametric tests of association (classic chi-square, Fisher's and Kruskal-Wallis) with a level of 5% (p-value < 0.05) being set for the rejection of the null hypothesis. RESULTS: Forty-seven over 18-year-old patients with sickle cell disease were evaluated. Most were black (78.7%) and female (59.6%) and the mean age was 30.1 years. The average number of bouts of pain annually was 7.02; pain was predominantly reported by individuals with sickle cell anemia (hemoglobin SS). The intensity of pain (Numeric Pain Scale) was 5.5 and the quantitative index (McGill) was 35.9. This study also shows that patients presented a high frequency of moderately painful crises in their own homes. CONCLUSION: According to these facts, it is essential that pain related to sickle cell disease is properly identified, quantified, characterized and treated at the three levels of healthcare. In primary healthcare, accurate measurement of pain combined with better care may decrease acute painful episodes and consequently minimize tissue damage, thus improving the patient's overall health.

Multiple sclerosis in South America: month of birth in different latitudes does not seem to interfere with the prevalence or progression of the disease.

OBJECTIVE: To assess whether the month of birth in different latitudes of South America might influence the presence or severity of multiple sclerosis (MS) later in life. METHODS: Neurologists in four South American countries working at MS units collected data on their patients' month of birth, gender, age, and disease progression. RESULTS: Analysis of data from 1207 MS patients and 1207 control subjects did not show any significant variation in the month of birth regarding the prevalence of MS in four latitude bands (0-10; 11-20; 21-30; and 31-40 degrees). There was no relationship between the month of birth and the severity of disease in each latitude band. CONCLUSION: The results from this study show that MS patients born to mothers who were pregnant at different Southern latitudes do not follow the seasonal pattern observed at high Northern latitudes.

Pain management at inpatient wards of a university hospital.

BACKGROUND AND OBJECTIVES: This is an exploratory, descriptive and transversal study aiming to determine the prevalence, characterization, location, and measurement and discuss pharmacological analgesic measures for acute pain management in five inpatient wards of a university hospital. METHOD: We enrolled 856 subjects in the study, of whom 272 were in pain at the time. Information related to pain was obtained using a bedside structured interview. Numeric pain scale and body diagram were used. RESULTS: Analgesia was assessed through medical records. The overall prevalence of pain was 31.8%, with severe pain in 44.2% and mean of 6.6 on numeric pain scale. The main reason was trauma and the most common site the abdomen. The most widely used analgesic was dipyrone (76.1%) with/without combination. Strong opioid was prescribed to 4.4%. For 27.5% there was no improvement. CONCLUSION: We conclude that pain is highly prevalent, poorly evaluated, undertreated, with inappropriate use of analgesics.

Clinical and epidemiological profile of patients with multiple sclerosis in Uberaba, Minas Gerais, Brazil.

UNLABELLED: Multiple sclerosis (MS) is an immune-mediated disease that affects the central nervous system. Clinical presentation and prevalence vary widely around the world. OBJECTIVE: To describe the clinical and epidemiological aspects of patients with MS in Uberaba (MG). METHOD: We conducted a transversal descriptive study, with data analysis of 35 patients with MS. RESULTS: Prevalence of MS was 12.5 cases/100,000 inhabitants, with a predominance in females (71.4%) and Caucasoid (85.7%). The current average age was (43.8 ys). The most common initial symptom was sensory (40%), followed by optical neuritis (25.7%). Expanded Disability Status Scale average score was 2.4. The relapsing-remitting form was predominant (88.6%), most (74.3%) were on immunomodulatory treatment and (40%) had college education. CONCLUSION: Prevalence of MS in Uberaba (MG) is considered average in accordance to Kurtzke and Page and clinical features are consistent with most Brazilian studies.

Overexpression of the plg1 gene encoding pectin lyase in Penicillium griseoroseum.

The pectin lyase (PL) is an industrially important enzyme since it is used for maceration and clarification in the process of fruit juice production in food industries. In order to increase the yields of pectin lyase we cloned the plg1 (pectin lyase 1) from Penicillium griseoroseum gene under the control of the strong constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) and the terminator region of the tryptophan synthetase (trpC) gene from Aspergillus nidulans (plasmid pAN52-Plg1) and transformed this construct into the P. griseoroseum strain PG63. One of the pAN52-Plg1 multi-copy transformants (strain 105) grown in culture medium containing glucose or sugar cane juice showed PL activities of 4,804 or 5,202 U ml(-1) respectively, which represented 57- and 132-fold increases. In addition, the apparent specific activity of PL produced by this strain was much higher than the one observed for a commercial pectinase preparation. Evaluation of the extracellular proteins in the culture supernatant of strain 105 by SDS-PAGE showed the presence of a clear and strong band of approximately 40 kDa that probably corresponds to PL. The enzyme yields reported here demonstrate that the system we developed is able to express pectin lyase at levels comparable to, or exceeding, previously reported data.

Sexuality and sickle cell anemia

BACKGROUND: Sickle cell disease, the most common hereditary blood disease in the world, is the result of an atypical hemoglobin called S (Hb S) which, when homozygous (Hb SS) is the cause of sickle cell anemia. Changes of puberty, correlated with a delayed growth spurt, begin late in both male and female sickle cell anemia individuals with repercussions on sexuality and reproduction. The objectives of this exploratory and descriptive study were to characterize the development of sexuality in adults with sickle cell anemia by investigating the patient's perception of their sex life, as well as the information they had and needed on this subject. METHODS: Twenty male and female sickle cell anemia patients treated at the Hemocentro Regional de Uberaba (UFTM) with ages between 19 and 47 years old were enrolled. A socioeconomic questionnaire and a semi-structured interview on sexuality, reproduction and genetic counseling were applied. RESULTS: This study shows that the sickle cell anemia patients lacked information on sexuality especially about the risks of pregnancy and the possible inheritance of the disease by their children. Moreover, the sexual life of the patients was impaired due to pain as well as discrimination and negative feelings experienced in close relationships. CONCLUSION: The health care of sickle cell anemia patients should take into account not only the clinical aspects of the disease, but also psychosocial aspects by providing counseling on sexuality, reproduction and genetics, in order to give this population the possibility of a better quality of life.

Pain measurement as part of primary healthcare of adult patients with sickle cell disease

OBJECTIVE: The aim of this exploratory, cross-sectional study was to evaluate pain in sickle cell disease patients and aspects related to primary healthcare. METHODS: Data were obtained through home interviews. The assessment instruments (body diagram, Numerical Pain Scale, McGill Pain Questionnaire) collected information on the underlying disease and on pain. Data were analyzed using the Statistical Package for Social Sciences program for Windows. Associations between the subgroups of sickle cell disease patients (hemoglobin SS, hemoglobin SC, sickle β-thalassemia and others) and pain were analyzed using contingency tables and non-parametric tests of association (classic chi-square, Fisher's and Kruskal-Wallis) with a level of 5% (p-value < 0.05) being set for the rejection of the null hypothesis. RESULTS: Forty-seven over 18-year-old patients with sickle cell disease were evaluated. Most were black (78.7%) and female (59.6%) and the mean age was 30.1 years. The average number of bouts of pain annually was 7.02; pain was predominantly reported by individuals with sickle cell anemia (hemoglobin SS). The intensity of pain (Numeric Pain Scale) was 5.5 and the quantitative index (McGill) was 35.9. This study also shows that patients presented a high frequency of moderately painful crises in their own homes. CONCLUSION: According to these facts, it is essential that pain related to sickle cell disease is properly identified, quantified, characterized ...

Multiple sclerosis in South America: month of birth in different latitudes does not seem to interfere with the prevalence or progression of the disease / Esclerose multipla na America do Sul: mes de nascimento em diferentes latitudes nao parece interferir com a prevalencia ou progressao da doenca

Objective To assess whether the month of birth in different latitudes of South America might influence the presence or severity of multiple sclerosis (MS) later in life. Methods Neurologists in four South American countries working at MS units collected data on their patients' month of birth, gender, age, and disease progression. Results Analysis of data from 1207 MS patients and 1207 control subjects did not show any significant variation in the month of birth regarding the prevalence of MS in four latitude bands (0–10; 11–20; 21–30; and 31–40 degrees). There was no relationship between the month of birth and the severity of disease in each latitude band. Conclusion The results from this study show that MS patients born to mothers who were pregnant at different Southern latitudes do not follow the seasonal pattern observed at high Northern latitudes. .

Objetivo Avaliar se o mês de nascimento em diferentes latitudes da América do Sul pode influenciar a presença ou gravidade da esclerose múltipla (EM) na vida. Método Neurologistas de quatro países da América do Sul trabalhando em unidades de EM coletaram os dados de seus pacientes com referência ao mês de nascimento, gênero, idade e progressão da doença. Resultados A análise dos dados mostrou que, para 1207 pacientes com EM e 1207 controles, não havia diferença significativa no mês de nascimento com relação à prevalência de EM em quatro zonas de latitude (0–10; 11–20; 21–30; e 31–40 graus). Não houve relação entre o mês de nascimento e a gravidade da doença em nenhuma destas zonas. Conclusão Os resultados deste estudo mostram que pacientes com EM nascidos de mães grávidas em diferentes latitudes sul não seguem o padrão dos resultados sazonais encontrados nas latitudes norte. .